Combination column

ABSTRACT

Combination columns comprised of individual connective parts, some of which may contain sieve means and other means, provide flexible and adaptive systems for effecting chemical processes such as column chromatography and involving carrier-bound reagents.

O United States Patent 1 [111 3,763,879

Jaworek Oct. 9,- 1973 COMBINATION COLUMN 3,360,128 12/1967 Federline210/232 x 3,522,884 8/1970 Frost 210/232 [75] Invenm" Upper 1,075,43310/1913 Milner 137/545 x Bavana, Germany 3,297,260 1/1967 Barlow 137/550x 984,082 2/1911 Edgerton 137/550 X [73] Ass'gnee' GmbH 2,819,799 1/1958Wilkerson 137/550 x y 868,022 10/1907 Shafer 137/550 x [22] m 19711,239,267 9/1917 Groves..... 137/544 x 3,407,841 10/1968 Semon 137/550 X[21] App1.No.: 116,581

Primary Examiner-Henry T. Klinksiek 1 Foreign Amfllcfllvn r ty DataAssistant Examiner-Robert J. Miller Feb. 23, 1970 Germany P 20 08 354.9Attorney-Burgess, Dinklage 8L Sprung [52] US. Cl 137/268, 210/232,210/266,

210/282, 137/550 [57] ABSTRACT 2 32 Combination columns comprised ofindividual con- 137/550 6 nective parts, some of which may contain sievemeans and other means, provide flexible and adaptive sys- [56]References Cited tems for effecting chemical processes such as columnUNITED STATES PATENTS chromatography and involving carrier-boundreagents. 1,927,228 9/1933 Compton 210/314 x 3 Claims, 7 Drawing Figuresv PATENTED BET 19 3 ShEF 1 UP 5 IN V EN TOR DIE TER ,JAWORE K BURGESS.DINKLAGE &SPRUNG AT TORNE Y5.

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DIE TER JAWOREK Pmgmmm 91315 7 3 79 IN VEN TOR DIE TE F? JAWORE KBURGESS. DINKLAGE & SPRUNG AT TORNE Y5 COMBINATION COLUMN The presentinvention is concerned with a combination, or composite, column which isespecially useful for a large variety of purposes in chemicaltechnology, especially in column chromatography and the like.

Due to recent developments in chemical technology, for example inchromatography, and in the use of carrier-bound reagents, such ascarrier-bound proteins and enzymes, the columns previously used haveproved to be too lacking in adaptability.

The present invention provides a combination column made up of certainindividual parts which can be connected together in any desired mannerand which, in the simplest manner, can be adapted to the most variedpossibilities of use and, in particular, permits the combination ofseveral chemical separation or reaction steps in a single column.

The combination column according to the present invention comprises atleast one cylindrical column segment provided with threads on both ends,as well as at least one separation piece which is also provided withthreads on both ends and in which a sieve is incorporated and/or atleast one end piece provided with a thread, the individual parts beingscrewed together in any desired manner.

All the parts of the combination column according to the presentinvention are preferably made from synthetic resin which is expedientlytransparent.

The column segments preferably have different lengths which bear adefinite relationship to one another. Thus, for example, in the case ofa column with a diameter of about 2 cm and comprising a plurality ofcolumn segments, column segments can be provided with lengths of l, 2, 4and 8 cm so that, by appropriate assembly of the individual columnsegments, it is possible to make up a column of any desired length.

The column segments preferably have an external thread on one end and acorresponding internal thread on the other end. However, in the case ofcolumns of small diameter, it is preferable that the threads on bothends are external, the-connection of the individual column segmentsbeing obtained by means of connection pieces provided with internalthreads at each end.

The separation pieces are similar to short column segments but contain asieve. The sieve is preferably set into the separationpiece in such amanner that it terminates one of the two ends of the separation piece.The sieve is also preferably inserted in such a manner that it can beexchanged so that sieves of different fineness can, as desired, beinserted.

The end pieces are preferably of massive construction and provided witha thread on at least one end with which they can be screwed onto acolumn segment or onto a separation piece. The end pieces are providedwith bores which can terminate externally in a nipple. This nipple canalso be constructed in such a manner that it can be screwed intoposition so that it can easily be removed and can, if desired, bereplaced by another nipple provided with a stopcock. The end of the endpiece provided with a thread is preferably countersunk somewhat in thedirection of the opening of the bore in order to promote a uniformrunning out of liquid from the column.

The combination column according to the present invention can also beprovided with massive intermediate pieces. A massive intermediate pieceof this type is provided with a thread on both ends and can thus bescrewed onto other column parts. It is provided with an axial bore whichis preferably narrow so that the volume of liquid which can be containedtherein is as small as possible. The bore can be provided with at leastone branch leading to the outside, this branch preferably terminating ina nipple which can be provided with a stopcock. Both ends of theintermediate piece are countersunk somewhat in the direction of the boreopening in the same manner as the above-mentioned one end of the endpiece.

According to one construction, the intermediate piece is provided withtwo branch bores and a four-way stopcock. In this way, it is possible tolead away to the outside, at certain points of the column, the liquidcontained therein either wholly or partially and to return it,optionally with addition liquid and/or reagents, into the followingcolumn segment.

The combination column according to the present invention is capable ofan extremely large number of variations. Thus, for example, by simplyscrewing together column segments and closure pieces, it is possible toconstruct columns of any desired but precisely defined length and ofprecisely defined column volume for any desired purpose. If, on the endsof a column assembled from one or more column segments, there arescrewed on separation pieces, then there is obtained a sieve cage openat both ends in which can be placed, for example carrier-bound proteins.The filled sieve cage can then be introduced into a liquid in the mannerof a tea egg for bringing about the corresponding reaction and can, whendesired, again be removed therefrom.

, When used in this manner, the combination column according to thepresent invention is expediently also provided with control pieces inthe form of a column segment, one end of which is cut at an angle andthe other straight end of which is threaded. The obliquely cut end ofthe control piece preferably has an angle of about 40 to 50 to thecylindrical axis. The thread of the straight end can be provided with acounter nut ring.

For carrying out a reaction with the use of a sieve cage of theabove-described type in a comparatively large volume of liquid, bothends of the sieve cage are preferably provided with control pieces. Oneof the control pieces, which is provided with a counter nut ring, isadjusted with regard to the other control piece in such a manner thatboth oblique openings are directed towards the same side. If, now, thecombination column constructed in this manner as a sieve cage" isrotated in a liquid, then the surrounding liquid flows through thecombination column and the desired reaction, for example an enzymaticreaction, can take place.

For the rotation of the column, there is preferably provided a magnetring which can be screwed on or off. This can consist of a fixedlyclampable ring which is pushed over the column and firmly clamped intoposition, this ring being provided with a stirrer magnet which, in theusual manner, rotates the column in conjunction with a magnetic stirrerpresent outside of the vessel.

The threads of the individual parts of the combination column accordingto the presetn invention are preferably provided with sealing rings inorder to prevent leakage of liquid when the column is in use.

A better understanding of the present invention and of severalembodiments thereof, may be had by refer ence to the accompanyingdrawings in which:

FIG. 1 is a perspective illustration of one column segment;

FIG. 2 is a perspective and exploded illustration of a separation piececomprising a sieve and sieve-holding means;

FIG. 3 is a perspective and exploded illustration of a combinationcolumn comprising three separate chambers and a T-piece; I

FIG. 4 is a perspective illustration of a combination column constructedas a sieve cage provided with control pieces and a magnet ring;

FIG. 5 is a partially sectional and perspective illustration of acombination column provided with a temperature regulating elementconsisting of two clamp rings and a column segment of comparativelylarge diameter;

FIG. 6 is another embodiment of a combination column with atemperature-regulating mantle; and

FIG. 7 is a further embodiment of a combination column with atemperature-regulating mantle.

More specifically, FIG. 1 shows a column segment 1 with an externalthread 10 and an internal thread lb. Instead of the internal thread lb,the column segment can also carry a second external thread la. In thelatter case, the individual column segments cannot be screwed togetherdirectly but require the help of short connecting pieces provided withappropriate threads.

FIG. 2 shows a separation piece 2 which has been broken up into itscomponent parts. The separation piece body 2 corresponds substantiallyto a short column segment and possesses an external thread 2a and aninternal thread at the other end (not illustrated). A sieve 3 placed inthe separation piece body 2 consists of the actual sieve piece 3a, whichis held by an inner ring 3b and an outer ring 3c and can be insertedinto one end of the body of separation piece 2 and is secured on a stopagainst axial displacement.

FIG. 3 shows the individual parts of a multi-chamber column according tothe present invention. Three column segments 1 of different length,together with three separation pieces 2 provided with sieves 3, two endpieces 4 and an intermediate piece 7, constitute a column with threeseparate chambers in which can be placed solid materials which may bethe same or different. Such solid materials can be, for example,exchangers, carrier-bound enzymes, molecular sieve substances orcombinations thereof. The end pieces 4 are provided with a thread 4a atone end and have a bore 5, the other end terminating in a nipple 6. Theinner end 4b of the end piece 4 is countersunk somewhat in the directionof the bore 5. The massive intermediate piece 7 has an axial bore 8. Abranching of the bore in the manner of a T-piece terminates in a nipple9. The intermediate piece 7 also has internal and external threads andthe two end surfaces thereof are countersunk somewhat-in the directionof the bore 8 in the same manner as in the end pieces 4.

FIG. 4 shows a combination column according to the present inventionassembled in the manner of a sieve cage. Both ends of a column segment 1carry separation pieces 2 (sieve 3 is not shown). The separation pieces2 carry, in turn, control pieces 10. By means of a counter nut ring 11,one of the control pieces is adjusted in such a manner that the obliqueouter end is brought in correspondence with the direction of the othercontrol piece. A synthetic resin ring 12, in which is mounted a stirrermagnet 13, is firmly clamped onto the column segment 1 by means of ascrew 14.

When the column combination illustrated in FIG. 4 is placed in a liquidcontainer which stands upon a magnetic stirrer, then the columncombination is caused to rotate by the magnet 13 and, by means of thecontrol pieces 10, in the case of suitable adjustment, a stream ofliquid is caused to pass through the column.

It will be readily appreciated that the combination columns according tothe present invention can be adapted to a large variety of differentuses. Thus, for example, column segments with separation pieces 2 can,when they are of suitable diameter, be placed in centrifuge glasses forcentrifuging off gels, the gel being retained by the sieve 3 but theliquid being centrifuged off. The desired depth of insertion into thecentrifuge beaker can be achieved, for example, by screwing up thecounter nut ring 11 on a column segment 1. Combined columns of the typeillustrated in FIG. 3 can be used for the sequential analysis ofproteins, for example, by placing identical or different enzymes boundto solid carriers in the column and passing the protein to be analyzedthrough the filled column. By the appropriate provision of separationpieces 3 or of intermediate pieces 7, it is possible to ascertain at anydesired point which amino acids or oligopeptides have been split off bythe continuing action of the proteolytic enzymes. Thus, the progress ofthe protein fission can be followed in a very simple manner without thefission itself being disturbed or inhibited in any way. Anotherpossibility is the combination of the reaction and the analysis in onecolumn; in a first column section, a particular reaction is allowed totake place and in a second column section a dissolved reagent isintroduced by means of an intermediate piece 7 so that a directdetermination can take place of the reaction occurring in column segment1.

Numerous other possibilities of using the combination column accordingto the present invention will be readily apparent. Thus, for example, aclamp ring can be provided which can be drawn over the column segmentsand separation pieces and clamping on of dialysis membranes, filterpapers, sieve pieces or the like thus made possible. From a columnsegment 1 and a ring 12 with magnet 13, there can thus be constructed amagnetically movable dialysis device in the simplest possible manner byclamping a dialysis membrane onto both ends. The clamping ring therebyconsists expediently of an inwardly open U-profile, the internaldiameter of which corresponds to the external diameter of the columnsegments and separation pieces, and of a rubber ring placed inside the Uwhich projects somewhat inwardly. The U-profile is made of a rigidmaterial, for example, of a synthetic resin.

According to a further embodiment, the clamping ring can have on atleast the outer end a thread suitable for screwing onto a further columnsegment of greater diameter and a bore which connects the ring outer endwith the ring end face bounding onto the thread so that, by means of twoclamping rings and a column segment of comparatively large diameter,there can be constructed a temperature-regulation mantle. A combinationcolumn with such a temperature-regulating mantle is illustrated in FIG.5. A combination column with column segments 1 and closure parts 4carries a heating or cooling mantle which consists of two clamping ringsl5 and a column segment 101 which has a greater diameter than the columnsegment 1. The cut-away clamping ring shows a rubber ring 16 placed inan inwardly open U-profile, which ring 16 firmly holds the clamping ringon the combination column and acts as a seal. The outside of theclamping ring 15 is provided with a thread 17 by means of which it isscrewed onto the column segment 101. A bore 18 connects the outside ofthe ring 19 with the ring end face 20 bordering onto the thread. Throughthe bores 18 there can be introduced and removed a cooling or heatingmedium, for example water, into and from the annular space between thecolumn segments 1 and 101. The openings on the outside of the ring 19 ofthe bores 18 are expediently provided with nipples.

FIG. 6 shows a further embodiment of a combination column according tothe present invention with a temperature-regulating mantle. In thiscase, the closure parts 4 are constructed in such a manner that theyhold the temperature-regulating mantle and permit the inflow and outflowof the temperature-regulating medium. The closure part 4 has a largerdiameter than the column segment 1 and, apart from thread 4a forscrewing onto the column segment 1, has a further thread 44 with alarger diameter which is suitable for screwing onto a column segment10]. In FIG. 6, this thread is illustrated as an internal thread but itcan also be constructed as an external thread. A bore 45 connects theFIG. 7 shows another embodiment of the combination column with atemperature-regulating mantle, this mantle being formed by thecombination of a closure part 4 as described in FIG. 6 with a clampingring according to FIG. 5 and a column segment 101. This Figure clearlyshows the many possibilities of alteration and adaptation which can beachieved by suitable combination of the constructional parts of thecolumn according to the present invention.

It will be understood that the specification and examples areillustrative but not limitative of the present invention and that otherembodiments within the spirit and scope of the invention will suggestthemselves to those skilled in the art.

What is claimed is:

1. Combination column for effecting chemical processes comprising atleast one hollow cylindrical column segment with threads on both ends,at least one hollow cylindrical threaded separating piece containingsieve means transversely disposed at one end thereof and at least onethreaded end piece, the said segments and pieces having a common axis.and being of substantially the same internal diameter and being screwedtogether in desired combination.

2. Column as claimed in claim 1 wherein there is more than one of saidcolumn segments and these column segments are of different length indefined length relationship to each other.

3. Column as claimed in claim 1 wherein said sieve means in saidcylindrical separating piece lies substantially in the same plane as oneof the ends of said separation piece.

1. Combination column for effecting chemical processes comprising atleast one hollow cylindrical column segment with threads on both ends,at least one hollow cylindrical threaded separating piece containingsieve means transversely disposed at one end thereof and at least onethreaded end piece, the said segments and pieces having a common axisand being of substantially the same internal diameter and being screwedtogether in desired combination.
 2. Column as claimed in claim 1 whereinthere is more than one of said column segments and these column segmentsare of different length in defined length relationship to each other. 3.Column as claimed in claim 1 wherein said sieve means in saidcylindrical separating piece lies substantially in the same plane as oneof the ends of said separation piece.